Molecular Epidemiological Survey Of Classical Swine Fever Virus In Guangxi And Preparation Of Monoclonal Antibodies

In this study, We investigate classical swine fever epidemic in Guang xi, collecting the CSFV suspected samples and nested RT-PCR method for detection, detection of CSFV nucleic acid samples。We select and Clone CSFV representative strain of E2. By Comprehensive analysis Combining the obtained sequence and pandemic strain sequence of CSFV in Guang xi downloading from GnenBank, it let us to comprehensive understanding of Guangxi the CSFV popular and genetic variation situation.From the results, We found Amino acid and nucleotide homology of the Guangxi pandemic strain、HCLV vaccine strains the and SHIMEN strains to change over time.the nucleotide and amino acids the highest decreased by3.7%.By constructing The phylogenetic tree of Guangxi CSFV pandemic strain and they belong to the two genes,4genotype,it contain1.1and2.1,2.2,2.3subsets gene.2.1subtype has become a popular strain of the advantages, no gene1.2subtypes and3-type strain. By Statistical data, conservation of pig disease still accounts for the first, but the piglets incidence rate showed an upward trend and increased to32.39percent from18.42percent in the past.1.1 subtype SHIMEN strains still exist in the Guangxi pigs, the main cause of sow reproductive failure and latent infection of newborn piglets incidence-based, gene2.1-type strain is mainly caused by the incidence of nursery pigs. for Further detecting of the CSFV antibody and found that latent infection of sow serum antibody positive (blocking rate≥40%) or negative (blocking rate<30%), We use nested PCR detection of nucleic acid positive.it indicate that Neutralizing antibodies after vaccination can not completely block the CSFV infection. this study investigate CSFV molecular epidemiological, to grasp the prevalence situation of Guang xi in recent years, classical swine fever virus, it provide basic data for the establishment of the Guangxi swine fever virus gene information database and the basis for the development of Guangxi swine fever prevention and control measures.In addition, the laboratory save and purified of classical swine fever viru immune BALB/c mice.we use monoclonal antibody technology was prepared by two secretion of anti-swine fever virus protein monoclonal antibody, they were named CSFV7and CSFV14.in the Monoclonal antibody identification,at the first,we found hybridoma culture supernatant titer of the CSFV7and CSFV14to1:256, ascites supernatant titer of CSFV7and CSFV14to1:106; at the second,they antibody subclass are IgG2b; at the third,we found relative affinity and index of CSFV7and CSFV14separate is1.5mol/L、1mol/L, that indicating They have the better affinity; at the last,by Western-blot analysis confirmed the IFA reaction of two monoclonal antibodies, monoclonal antibody not only specific with CSFV response, and they against recombinant E2proteins of CSFV. Preparation Monoclonal antibody for further rapid detection of classical swine fever virus antigen lay the foundation...